Ultrasensitive dual-color rapid lateral flow immunoassay via gold nanoparticles with two different morphologies for the serodiagnosis of human brucellosis.

Abstract

Lateral flow immunoassays (LFIAs) are popular because they are rapid, convenient, stable, low cost, and easy to read. However, conventional LFIAs based on gold nanoparticles lack sensitivity, which hinders their widespread use. Here, we prepared durian-like gold nanoparticles (GNDs) and labeled them with staphylococcal protein A to detect brucella antibody. Then, the analytical performances of GNDs and gold nanospheres (GNSs) with the same diameter were compared. It was found that the sensitivity of GNDs was five to ten times higher than that of GNSs. The nonspherical morphologies of the nanoparticles greatly increased the sensitivity of the LFIA. On the basis of GNDs and GNSs, we developed an ultrasensitive dual-color brucellosis LFIA. GNSs labeled with streptavidin were used to demonstrate the control line. This dual-color LFIA had a diagnostic sensitivity and specificity of 100%. Human standard Brucella-positive serum (containing brucella antibody at 4000 IU/mL) could be detected in this system even for a dilution factor of 10-5. The detection limit was 0.04 IU/mL. This is two orders of magnitude better than conventional LFIA strips (detection limit 4 IU/mL). This dual-color LFIA contains all components of a conventional LFIA with no additional processing steps or reagents. It can detect antibodies in serum, plasma, and even whole blood without sample pretreatment or blood filtration pads. Both types of nanoparticles were synthesized in a simple and low-cost manner. This suggests that it will have utility for the early diagnosis of brucellosis and other diseases. Graphical abstract.